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Development of Cross-Species EST-SSR markers based on Transcriptome Sequence analysis of Morinda officinalis(Rubiaceae)

Boyong Liao  Kaikai Meng  Qianyi Yin  Cuiying Huang  Hanzhang Song  Qiang Fan  Wenbo Liao  Sufang Chen  
【摘要】:Morinda L.(Rubiaceae) contains approximately 80-100 species,many of which are famous local medical plants.However,the survival of the natural populations of these species are always threatened by excessive collections,calling for widely scientific attentions.M.officinalis is,known as one the four medicine plants in South China,becomes extinct in the natural environments.In this study,we present the first transcriptome analysis of M.officinalis using the Illumina sequencing technology.More than 11 million raw reads were generated,and de novo assembled into 51,049 unigenes,with an average length of 801 bp.The similarity search indicated that 25,547 unigenes(50.04 %) had significant similarity with proteins in the NCBI Non-redundant(NR) database,25,546 and 18646 unigenes were assigned with Gene Ontologies(GO) and Eukaryotic Orthologous Groups(KOG),respectively,and a total of 9,886 unigenes were mapped into 241 pathways in the Kyoto Encyclopedia of Genes and Genomes Pathway database(KEGG).Some abundant transcripts related to Genetic information,including Folding,sorting and degradation(843,8.53%) and Translation(770,7.79%) were identified in the M.officinalis transcriptome.In total,8,064 potential EST-SSRs were identified among the unigenes.The most abundant type of repeat was mononucleotide(4221,52.34 %),followed by trinucleotide(1588,19.69 %) and dinucleotide(1040,12.90 %) repeats in perfect EST-SSRs.Finally,5605 SSR primer pairs were designed successful.In addition,a total of 100 potential marker sites were selected to validate the assembly quality,and 24 pairs of polymorphic primers were characterized using four populations of M.officinalis from Guangdong province.This study generated a substantial fraction of M.officinalis transcriptome sequences,which were very useful resources for gene discovery and molecular marker development.

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